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A method for enriching cysteine-containing peptides based on fluorine solid phase extraction and performing mass spectrometry analysis

A cystine peptide, extraction and enrichment technology, applied in the field of separation and enrichment of cysteine ​​peptides in proteolytic peptides, can solve the problem of high cost, limited popularization of fluorine solid phase extraction technology, and no graphite fluoride yet. materials and other issues, to achieve the effect of low cost, good fluorine solid phase extraction performance, and good enrichment effect

Active Publication Date: 2021-02-26
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that with the development of organic fluorine chemistry, there are a wide variety of derivative reagents with fluorine labels, which greatly promotes the development of fluorine solid-phase extraction technology. As a matrix, it is made by bonding a highly fluorinated silane coupling agent, and its cost is relatively high, which limits the further popularization of fluorine solid phase extraction technology
Fluorinated graphite is a kind of cheap and easy-to-obtain high-fluorinated materials, which are often used as solid-phase lubricants and lithium battery cathode materials in industry, but fluorinated graphite materials have not been used as solid-phase adsorption for fluorine solid-phase extraction. agent reports

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  • A method for enriching cysteine-containing peptides based on fluorine solid phase extraction and performing mass spectrometry analysis
  • A method for enriching cysteine-containing peptides based on fluorine solid phase extraction and performing mass spectrometry analysis
  • A method for enriching cysteine-containing peptides based on fluorine solid phase extraction and performing mass spectrometry analysis

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Effect test

Embodiment 1

[0026] Graphite fluoride as fluorine solid-phase extraction adsorbent for enrichment selectivity investigation standard cysteine-containing peptide (amino acid sequence: AGVHGCNGLR) 1 microgram and equine myoglobin (Myoglobin) enzymatic peptide with 1: After mixing at 100 (molar ratio), in 100 microliters of 50% acetonitrile, 25 mmol / L ammonium bicarbonate aqueous solution, carry out fluorine-labeled derivatization at 50 degrees Celsius for 2 hours under dark conditions (derivatization reagent: N-[(3- Perfluorooctyl)propyl]iodoacetamide) and freeze-dried, the peptide mixture was redissolved in 50% methanol and 25 mmol / L ammonium bicarbonate aqueous solution, added 4 mg of graphite fluoride, and carried out fluorine solid-phase extraction. The solution before and after elution and enrichment was analyzed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS), and the obtained spectra were as follows: figure 2 As shown, the results show t...

Embodiment 2

[0028] Graphite fluoride used as fluorine solid-phase extraction adsorbent for enrichment detection limit inspection standard cysteine-containing peptide (amino acid sequence: AGVHGCNGLR) derivatized with fluorine reagent (derivatization reagent: N-[(3-Perfluorooctyl)propyl ]iodoacetamide), take a certain amount, redissolve in 50% methanol, 25 mmol / L ammonium bicarbonate aqueous solution, respectively, to obtain 100 nmol / L and 10 nmol / L solutions, add 4 mg of graphite fluoride, and carry out fluorine solidification phase extraction, and the enriched solution was analyzed by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). image 3 As shown, the results show that graphite fluoride has a lower detection limit as a fluorine solid phase extraction material.

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Abstract

The invention belongs to the field of analytical chemistry and protein analysis, relates to a new method of fluorine solid-phase extraction, in particular to a method for separating and enriching cysteine-containing peptides by using fluorinated graphite as a solid-phase adsorbent. The present invention uses fluorinated graphite as a solid-phase adsorbent for fluorine solid-phase extraction, and utilizes the fluorine-fluorine interaction between the surface of the fluorinated graphite and the fluorine label of the target analyte to effectively separate the target analyte and impurities. Compared with the traditional fluorine solid-phase extraction adsorbent, the fluorinated graphite used in the present invention avoids the use of high-purity silica matrix and high-fluorinated silane coupling agent, and has low cost, good selectivity, and high detection efficiency. With the characteristics of low limit, high recovery rate and high popularity, it can realize the separation, enrichment and mass spectrometry identification of cysteine-containing peptides.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and protein analysis, and relates to a new method of fluorine solid-phase extraction, in particular to a fluorine solid-phase extraction technology using graphite fluoride as a solid-phase adsorbent, and using the technology to separate and enrich proteolytic peptides Method for cysteine ​​peptides. Background technique [0002] The prior art discloses that fluorine solid phase extraction is a widely used technique in organic synthesis and catalysis. This technology derivatizes the target compound with a fluorine reagent, utilizes fluorine-fluorine interaction, and relies on highly fluorinated solid-phase adsorbents to separate and purify the target compound. Practice has shown that compared with traditional biotin tags, fluorine tags are more compatible with mass spectrometry, so in recent years it has been used in various "omics" research (such as proteomics, glycomics, metabolomics) have b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848
Inventor 陆豪杰张程张莹
Owner FUDAN UNIV